Czech Academy of Sciences, Institute of Analytical Chemistry, Veveří 967/97, Brno 602 00, Czech Republic; Department of Chemistry, Faculty of Science, Masaryk University, Kamenice 5, Brno 625 00, Czech Republic.
Czech Academy of Sciences, Institute of Analytical Chemistry, Veveří 967/97, Brno 602 00, Czech Republic.
J Chromatogr A. 2022 Dec 6;1685:463591. doi: 10.1016/j.chroma.2022.463591. Epub 2022 Oct 20.
With increasing demands on protein analyses in complex biological matrices, the insistence on developing new sample preparation techniques is rising. Recently, we introduced a new displacement electrophoresis technique (epitachophoresis) and instrumentation for preparative concentration and cleaning of DNA samples. This work describes the possibility of applying this device to protein samples. We have developed a method for the epitachophoretic concentration of proteins in a cationic mode and tested it by concentrating and collecting the protein zones from complex biological matrices (urine and growth medium). Under optimized conditions, we have obtained recoveries up to 99%. Furthermore, the applicability of the developed method was proven by concentrating and collecting the cytochrome c zone from a HeLa cell line growth medium, where the protein cytochrome c was released during cell apoptosis.
随着对复杂生物基质中蛋白质分析的需求不断增加，开发新的样品制备技术的要求也在不断提高。最近，我们引入了一种新的电泳置换技术（电泳置换电泳）和仪器，用于 DNA 样品的制备浓缩和清洗。本工作描述了将该装置应用于蛋白质样品的可能性。我们开发了一种在阳离子模式下电泳浓缩蛋白质的方法，并通过浓缩和收集来自复杂生物基质（尿液和生长培养基）的蛋白质区带对其进行了测试。在优化条件下，我们获得了高达 99%的回收率。此外，通过从 HeLa 细胞系生长培养基中浓缩和收集细胞色素 c 区带，证明了所开发方法的适用性，其中在细胞凋亡过程中释放了蛋白质细胞色素 c。
